Method for treating and preventing inflammation and atherosclerosis

ABSTRACT

This invention provides a method for treating or preventing inflammation or atherosclerosis in mammals comprising administering an effective amount of a 15-LO inhibitor of Formula I: ##STR1## wherein: R 1  is hydrogen or C 1  -C 6  alkyl; 
     R 2 , R 3 , R 4 , and R 5  independently are hydrogen, C 1  -C 6  alkyl, nitro, halo, CN, OR 6 , NR 6  R 7 , --CO 2  R 6 , CONR 6  R 7 , CH 2  OR 6 , or CH 2  NR 6  R 7 , and R 2  and R 3 , and R 4  and R 5 , when attached to adjacent ring atoms, can be --(CH 2 ) 3  or 4 --; 
     in which R 6  and R 7  independently are hydrogen, C 1  -C 6  alkyl, phenyl or benzyl, and when taken together with the nitrogen to which they are attached, R 6  and R 7  can complete a cyclic ring having from 3 to 7 carbon atoms; ##STR2## in which R 8 , R 8&#39; , R 9 , and R 9&#39;   independently are hydrogen or C 1  -C 6  alkyl, n is 0, 1, or 2, and Z.sup.⊖ is an anion, and pharmaceutically acceptable salts thereof.

This application is a 371 of PCT/US96/14242 filed Sept. 5, 1996.

This invention concerns a method for treating and preventinginflammation or atherosclerosis in mammals by administering a compoundwhich is an inhibitor of the enzyme 15-lipoxygenase (15-LO).

BACKGROUND OF THE INVENTION

Atherosclerosis is a multifactorial disease characterized by excessiveintracellular lipid deposition in macrophages, leading to formation offoam cells. The accumulation of lipid-loaded foam cells in thesubendothelial space leads to formation of fatty streaks, which are theearly atherosclerotic lesions. Oxidative modification of lipids,specifically low-density lipoprotein, has been implicated as a majorprocess in foam-cell formation.

Lipoxygenases are nonheme iron-containing enzymes that catalyze theoxygenation of certain polyunsaturated fatty acids such as lipoproteins.Several different lipoxygenase enzymes are known, each having acharacteristic oxidation action. One specific lipoxygenase, namely15-LO, has been detected in atherosclerotic lesions in mammals,specifically rabbit and man. The enzyme, in addition to its role inoxidative modification of lipoproteins, is important in the inflammatoryreaction in the atherosclerotic lesion. Indeed, 15-LO has been shown tobe induced in human monocytes by the cytokin IL-4, which is known to beimplicated in the inflammatory process.

Another class of lipoxygenase enzymes is 5-lipoxygenase (5-LO). Whilethis enzyme causes oxidation of unsaturated fatty acids, it primarily isresponsible for inserting oxygen on position 5 of arachidonic acid.Other lipoxygenases are known; one of the most common and abundant being12-lipoxygenase (12-LO).

We have now found that inhibitors of 15-LO are especially useful toprevent and treat inflammation and atherosclerosis. While there areseveral lipoxygenase enzymes, specific inhibition of 15-LO is criticalin the inflammatory and atherosclerosis process. All that is requiredaccording to this invention is to administer a 15-LO inhibitor, andespecially one that is a specific 15-LO inhibitor.

Several classes of organic compounds are 15-LO inhibitors.

Tetracyclic indole and benzopyranoindole compounds are potent 15-LOinhibitors. U.S. Pat. No. 3,388,133 describesbenz[b]indolo[2,3-d]-thiopyrano and pyrylium salts as antibacterial andantifungal agents. U.S. Pat. No. 4,132,714 describes a process formaking chromenoindoles, which are said to be useful as color-formingagents. U.S. Pat. No. 4,797,495 discloses a wide variety ofbenzocarbazoles which are said to be antitumor agents. Similarly,benzimidazoles are well known as antiviral agents. U.S. Pat. No.4,293,558 describes various 1-thiazolinyl-2-aminobenzimidazoles, andU.S. Pat. No. 4,243,813 describes 1-sulfonyl-benzimidazoles. None ofthose compounds have been described as inhibitors of 15-LO, and nonehave been utilized in treating inflammation or atherosclerosis. All ofthese compounds are 15-LO inhibitors and can be employed in the methodof this invention.

We have now discovered that compounds which are effective inhibitors of15-LO are useful in treating and preventing inflammation andatherosclerosis.

SUMMARY OF THE INVENTION

This invention provides a method for treating and preventinginflammation or atherosclerosis in mammals comprising administering aneffective amount of a 15-LO inhibitor. The invention preferably employsa specific 15-LO inhibitor. In a preferred embodiment, the 15-LOinhibitor is a benzopyranoindole or related compound as described inU.S. Pat. Nos. 3,388,133, 4,132,714, and 4,797,495, which areincorporated herein by reference. Especially preferred 15-LO inhibitorshave Formula I ##STR3## wherein: R¹ is hydrogen or C₁ -C₆ alkyl;

R², R³, R⁴, and R⁵ independently are hydrogen, C₁ -C₆ alkyl, nitro,halo, CN, OR⁶, NR⁶ R⁷, --CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or CH₂ NR⁶ R⁷, andR² and R³, and R⁴ and R⁵, when attached to adjacent ring atoms, can be--(CH₂)₃ or 4 --;

in which R⁶ and R⁷ independently are hydrogen, C₁ -C₆ alkyl, phenyl orbenzyl, and when taken together with the nitrogen to which they areattached, R⁶ and R⁷ can complete a cyclic ring having from 3 to 7 carbonatoms; ##STR4## in which R⁸, R^(8'), R⁹, and R^(9') independently arehydrogen or C₁ -C₆ alkyl, n is 0, 1, or 2, and Z.sup.⊖ is an anion, andpharmaceutically acceptable salts thereof.

A preferred method according to this invention employs a compound of theformula ##STR5## where R¹, R², R³, R⁴, R⁵, R⁸, and Z.sup.⊖ have theabove defined meanings. Within this group, preferred compounds to beemployed are those wherein R¹ is hydrogen, and one or two of R², R³, R⁴,and R⁵ are selected from C₁ -C₆ alkyl, halo, nitro, or OR⁶, where R⁶ ispreferably C₁ -C₆ alkyl.

Another preferred embodiment utilizes compounds of the formula ##STR6##where R¹, R², R³, R⁴, R⁵, R⁸, and R^(8') are as defined above.

Another preferred method of treatment employs a compound of the formula##STR7## where R¹, R², R³, R⁴, R⁵, R⁸, and R⁹ are as defined above.

Benzimidazole 15-LO inhibitors to be employed in the method of thisinvention are known and readily available as described in any of thefollowing United States patents, all of which are incorporated herein byreference: U.S. Pat. Nos. 3,853,908; 3,682,952; 3,850,954; 4,118,742;4,196,125; 4,216,313; and 4,492,708. Additional benzimidazoles aredescribed in the book entitled Benzimidazoles and Congeneric TricyclicCompounds, P. N. Preston, Ed., John Wiley & Sons, also incorporatedherein by reference.

In a preferred embodiment, the 15-LO inhibitor utilized is abenzimidazole having the Formula II ##STR8## where R² and R³independently are hydrogen, C₁ -C₆ alkyl, nitro, halo, CN, OR⁶, NR⁶ R⁷,--CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or CH₂ NR⁶ R⁷, and R² and R³ when attachedto adjacent ring atoms can be --(CH₂)₃ or 4--;

R⁶ and R⁷ are as defined above;

R¹⁰ is SO₂ R¹², hydrogen, C₁ -C₆ alkyl, phenyl, or phenyl substitutedwith 1, 2, or 3 groups selected from halo, CN, OR⁶, C₁ -C₆ alkyl, NR⁶R⁷, CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or CH₂ NR⁶ R⁷ ; and

R¹¹ and R¹² independently are hydrogen, halo, NR⁶ R⁷, OR⁶, C₁ -C₆ alkyl,C₃ -C₇ cycloalkyl optionally containing an O, N, or S atom, phenyl, orphenyl substituted by 1, 2, or 3 groups selected from halo, CN, OR⁶, C₁-C₆ alkyl, NR⁶ R⁷, --CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or CH₂ NR⁶ R⁷.

Another preferred method employs a benzimidazole of the formula ##STR9##where R¹³ is phenyl, 4-chlorophenyl, 4-fluorophenyl, 4-nitrophenyl,2,5-dichlorophenyl, 2-furanyl, 2-thienyl, 3-pyridyl, or 4-pyridyl.

In another embodiment, the 15-LO inhibitor utilized is a substitutedindole. Typical indoles which can be employed include the carbamates andureas of Formula III ##STR10## wherein Y is CH₂, S, O, or NR⁷, and R¹,R², R³, and R⁶ are as defined above, and Ar is phenyl, Het, and phenylor Het substituted with 1, 2, or 3 groups selected from halo, CN, OR⁶,C₁ -C₆ alkyl, NR⁶ R⁷, --CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or CH₂ NR⁶ R⁷, whereHet is a heterocyclic group selected from thiophene, furan, pyrrole,isopyrrole, pyrazole, 1,2,3-triazole, 1,2,4-triazole, oxazole,isoxazole, thiazole, isothiazole, 1,2,3-oxadiazole, thiazine, pyridine,pyrazine, piperazine, pyrrolidine, piperidine, and pyridazine, and whereHet is optionally substituted with phenyl or substituted phenyl,furanyl, thienyl, or pyridyl, and where R⁶ and R⁷ are as defined above.A particularly preferred group of compounds to be employed in the methodhave the formula ##STR11## where R¹, R², R³, R⁴, R⁵, R⁶, and Y are asdefined above. Such compounds are described in EP 0150505, which isincorporated herein by reference.

Another class of 15-LO inhibitors which can be utilized in the inventionare styrenes having the Formula IV ##STR12## wherein Ar and Ar'independently are phenyl, Het, and phenyl or Het substituted with 1, 2,or 3 groups selected from halo, OR⁶, C₁ -C₆ alkyl, NR⁶ R⁷, --CO₂ R⁶ ₁,CONR⁶ R⁷, CH₂ OR⁶, and CH₂ NR⁶ R⁷, where R⁶ and R⁷ are as defined above.

A preferred method employs styrene 15-LO inhibitors of the formula##STR13## where R², R³, and Het are as defined above.

Another group of 15-LO inhibitors are catacholes, compounds of theFormula V ##STR14## wherein R², R³, and R⁶ are as defined above.

Still other 15-LO inhibitors that can be utilized are naphthalenes,especially those of Formula VI ##STR15## wherein R¹, R², R³, and R⁶ areas defined above, and M is hydrogen or a cation such as sodium,potassium, or calcium. Such compounds are described in U.S. Pat. No.4,608,390, incorporated herein by reference.

Another class of 15-LO inhibitors are benzoxadiazines of the generalFormula VII ##STR16## where R¹, R², R³, and Ar are as defined above.Such compounds are described in EP 0410834.

A preferred embodiment utilizes a benzo[a]phenothiazine which isdescribed in U.S. Pat. No. 4,876,246, incorporated herein by reference.Related 15-LO inhibitors that can be employed are phenothiazonederivatives described in U.S. Pat. No. 4,939,145, incorporated herein byreference.

Especially preferred from such classes are compounds having the formulas##STR17## where R¹, R², R³, R⁴, R⁵, and Y are as defined above. Suchcompounds are specifically described in U.S. Pat. Nos. 4,876,246 and4,939,145.

All that is required to practice the method of this invention is toadminister to a mammal a 15-LO inhibiting amount of a 15-LO inhibitor,preferably a specific 15-LO inhibitor.

DETAILED DESCRIPTION OF THE INVENTION

The term "C₁ -C₆ alkyl" means a straight or branched carbon chain suchas methyl, ethyl, isopropyl, n-butyl, tert-butyl, sec.-pentyl,3-methylpentyl, and the like. "Halo" means fluoro, chloro, bromo, andiodo. Ring substituents R², R³, R⁴, and R⁵ can be OR⁶, where R⁶ can behydrogen or C₁ -C₆ alkyl. Typical groups defined by OR⁶ include hydroxy,methoxy, isopropoxy, tert-butoxy, n-hexyloxy, and the like. Ringsubstituents also are defined by NR⁶ R⁷, which groups include amino,methylamino, diethylamino, N-methyl-N-isohexylamino, and the like. Thering substituents R², R³, R⁴, and R⁵ can also be a carboxylic acid,ester, carboxamide, and methylamino group. Typical esters includemethoxycarbonyl and ethoxycarbonyl. Typical carboxamide groups includeaminocarbonyl, methylamino-carbonyl and N,N-diethylaminocarbonyl.Typical methylamino groups include methylaminomethyl, ethylaminomethyl,and the like.

The term "Z.sup.⊖ " in the above formula is an anion such as perchlorateor halide, for instance chloride, bromide, or the like.

The compounds to be employed in the method of this invention are known.They can be prepared by processes described in the art. For example,U.S. Pat. No. 3,388,133, which is incorporated herein by reference,describes reaction of a phenylhydrazine with a thiochroman-4-one to givecompounds of Formula I wherein X is ##STR18## This reaction scheme isapplicable to other compounds, for example according to the followingscheme ##STR19##

The 15-LO inhibitors are effective for treating inflammation andatherosclerosis. A characteristic feature of atherosclerosis is theaccumulation of cholesterol ester engorged from foam cells. Foam cellsare derived from circulating monocytes which invade artery walls inresponse to hypercholesterolemia, and mature into tissue macrophages.The enzyme 15-LO has been implicated in inflammatory disorders and inthe origin and recruitment of foam cells (see Harats, et al., TrendsCardioivasc. Med., 1995;5(1):29-36). This enzyme is capable of oxidizingesterified polyenoic fatty acids, such as those found in phospholipids.Treatment of experimental animals with antioxidants which reducehydroperoxides produced by 15-LO has been shown to retard theprogression of atherosclerotic lesions. Accordingly, administeringcompounds which inhibit 15-LO is an effective way to treat and preventatherosclerosis.

The compounds described above are effective inhibitors of 15-LO whenevaluated in standard assays routinely utilized to measure 15-LOactivity. Specifically, representative compounds were evaluated by themethods described by Auerbach, et al., Analytical Biochemistry,1992;201:375-380. Two in vitro assays were utilized, both utilizingrabbit reticulocyte 15-LO, and linoleic acid as substrate, toenzymatically produce a peroxide oxidation product known as 13(S)-HPODE.N-Benzoyl leucomethylene blue was utilized as a calorimetric reagent fordetection and quantification of the peroxide formation. Also, HPLC wasutilized to quantify the oxidation following incubation at 4° C. for 10minutes.

The 15-LO inhibitory activity of representative compounds is presentedin Table 1. Data Column 1 gives the concentration of compound requiredto inhibit 50% of the activity of 15-LO (IC₅₀) when measured by the HPLCmethod of Auerbach, et al. Data Column 2 gives the concentration ofselected compounds to inhibit 50% of the activity of the 5-LO enzyme.

                  TABLE 1                                                         ______________________________________                                        Compounds of the Formula                                                        #STR20##                                                                       -                                                                                                              15-LO                                       R.sup.2 R.sup.3 R.sup.4 R.sup.5 R.sup.8 IC.sub.50 μM 5-LO                ______________________________________                                          H H H H H 1.0-4.0                                                             H H NO.sub.2 H H 0.48 >10                                                     H H Cl H H 12.0                                                               H H CH.sub.3 H H 1.9                                                          H H H CH.sub.3 H 1.0-4.0                                                    H       H        --(CH.sub.2).sub.4 --                                                                    H     0.5-2.0                                                                              >32                                  H       CH.sub.3 O--                                                                           H       H    H     0.70   >9.6                                 CH.sub.3 O H H H H 26.0                                                       H H H H CH.sub.3 >25                                                          #STR21##                                                                      H         H        H     H    H     3.8    22                                 #STR22##                                                                      H         H        H     H    H     1.3    >100                               H CH.sub.3 O-- H H H 0.6 >10                                                  CH.sub.3 O-- H H H H 4.5 >50                                                  #STR23##                                                                      H         CH.sub.3 O--                                                                           H     H    H     4.0    >50                                #STR24##                                                                      H         CH.sub.3 O--                                                                           H     H    H     >25 μm                                 #STR25##                                                                      H         CH.sub.3 O--                                                                           H     H    H     >25 μM                                 #STR26##                                                                      H         H        H     H    H     1.5    >10                              ______________________________________                                    

As noted above, benzimidazoles are especially preferred 15-LO inhibitorsto be employed in the claimed method. The 15-LO inhibitory activity oftypical benzimidazoles are given in Table 2.

                  TABLE 2                                                         ______________________________________                                        Compounds of the Formula                                                        #STR27##                                                                       -                                                                                                            15-LO  5-LO                                   R.sup.2 R.sup.3 R.sup.10 R.sup.11 IC.sub.50 μM IC.sub.50 μM           ______________________________________                                          H H H                                                                                                                  1.50 8##                                                                    >10                                     - H 5-Cl H                                                                                                            0.65 >10                              - H 5-Cl H                                                                                                            0.7 >10                               - H 5-Cl H                                                                                                            0.24 >10                           ______________________________________                                    

Styrenes are potent 15-LO inhibitors which can be employed in thepresent method. Table 3 gives the 15-LO inhibitor of typical styrenes.

                  TABLE 3                                                         ______________________________________                                          #STR32##                                                                       -                            15-LO 5-LO                                      R.sup.2 R.sup.3 Het IC.sub.50 μM IC.sub.50 μM                         ______________________________________                                          H 4-OCH.sub.3                                                                                                     1.6 >10#                                ______________________________________                                    

Typical indole 15-LO inhibitors which can be utilized have theactivities shown in Table 4.

                  TABLE 4                                                         ______________________________________                                          #STR34##                                                                       -                                      15-LO  5-LO                           R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5 R.sup.6 Y IC.sub.50 μM                                                            IC.sub.50 μM              ______________________________________                                        H   H      H      H    3-Cl  H    S     4      >10                              H H H H H H CH.sub.2 1.5 >10                                                ______________________________________                                    

Table 5 gives additional selectivity data for typical 15-LO inhibitorswhich can be utilized in the method of this invention.

                  TABLE 5                                                         ______________________________________                                                             15-LO  5-LO                                                IC.sub.50 μM IC.sub.50 μM                                             ______________________________________                                                                          2.0 35##                                                                    >30                                              -                                                                                                            1.5 >10                                        - CH.sub.3 (CH.sub.2).sub.3 (CH.sub.2 --C.tbd.C--).sub.4 (CH.sub.2).sub                                    .3 --COOH 0.75 >10                            ______________________________________                                    

As further evidence of 15-LO inhibitors being effective to prevent andtreat inflammation and atherosclerosis in animals, one representativecompound has been extensively evaluated in cholesterol-fed rabbits overa 12-week period. The compound evaluated "Compound A" was6,11-dihydro[1]benzothiopyran[4,3-6]-indole, the compound of Formula Iwhere R¹, R², R³, R⁴, and R⁵ each are hydrogen, and X is --CH₂ --S--,i.e., ##STR37##

Specific pathogen-free New Zealand White rabbits (≈2.5 kg) were obtainedfrom Myrtle's Rabbitry (Thompson Station, Tenn.). The animals were fed astandard laboratory diet (Ralston Purina, St. Louis, Mo.) and wereallowed to become acclimatized for 7 days before initiation of thestudy, at which time two groups of rabbits (n=10 each) were begun on adiet enriched with cholesterol (0.25% wt/wt), peanut oil (3% wt/wt), andcoconut oil (3% wt/wt), with a small amount of applesauce mixed into thefood to enhance palatability. This diet was designed to produce a modesthypercholesterolemic response. The control group received no additionaltreatment. The drug-treated group received 350 mg of Compound A perkilogram body weight per day in their food. Rabbits were permittedaccess to 40 g of food at ≈12 hour intervals via automated feeders, anddiet intake was monitored every day such that the animals received 175mg/kg/bid. Water was available ad libitum. Body weights were measured atregular intervals throughout the 12-week study. Blood samples wereobtained at the indicated intervals for determination of hematocrit andplasma lipid concentrations.

Characterization of Atherosclerotic Lesions

Rabbits were euthanized by an overdose of sodium pentobarbital (150mg/kg⁻¹) and exsanguinated via the abdominal aorta. Aortas were removedfrom the valve to the ileal bifurcation, opened to expose the intima,and photographed with a Polaroid camera. By use of these photographs,the areas of grossly discernible atherosclerosis were manuallyintegrated on a digitizing pad and calculated with SigmaScan (JandelScientific). Aortas were visually subdivided into three areas: arch(aortic valve to first intercostal), thoracic aorta (first intercostalto diaphragm area), and abdominal aorta (diaphragm to ilealbifurcation). In addition to extracting aortas, body tissues weresurveyed for indications of adverse reactions.

Determination of Cholesterol Esters and Unesterified Cholesterol Content

Weighed segments of each aortic region (arch, thoracic, and abdominal)were extracted. Esterified and unesterified cholesterol content ofaortic tissue were determined by gas chromatography using 5-α-cholestaneas an internal standard.

In the control group, the arch area of aortic sections demonstratedabout 15% lesion coverage of intima, whereas those animals receivingCompound A showed no lesion coverage. No detectable lesions were seen ineither group in the thoracic region. In the abdominal region, thecontrol group exhibited 5% lesion coverage, whereas the treated groupexhibited about 1%. The treated group had no detectable cholesterolesters present in the arch, thoracic, or abdominal regions, whereas thecontrol group had about 2 mg/g tissue wet weight of cholesterol estersin the arch region, none in the thoracic region, and about 0.6 mg/g inthe abdominal region. Test animals and the control group had about thesame amount of unesterified cholesterol in the thoracic and abdominalregions (0.7-0.8 mg/g tissue wet weight), while in the arch region, thecontrol group had about 1.4 mg/g while the treated group had about 0.8mg/g.

These data establish that administration of a 15-LO inhibitoreffectively protects against the development of atherosclerosis inanimals.

In an especially preferred embodiment of this invention, the 15-LOinhibitor utilized is a specific inhibitor of 15-LO. The term "specific"as used herein means that a compound inhibits 15-LO at least aboutten-fold (10×) more effectively than it inhibits 5-LO. For example, apreferred group of compounds to be employed in the present method aredefined by Formula I. A typical compound from within that group is6,11-dihydro[1]benzothiopyrano[4,3-b]-indole (Compound A). Its 15-LOinhibitory activity is an IC₅₀ of 1.3 μM, and its 5-LO inhibitoryactivity is >100 μM. The compound thus inhibits 15-LO at least about 100times more potently than it inhibits 5-LO. The compound is therefore a"specific" 15-LO inhibitor for purposes of this invention.

Similarly, a preferred benzimidazole to be employed in the invention is2-(4-chlorophenyl)-5-chlorobenzimidazole. It has a 15-LO IC₅₀ of 0.65μM, and a 5-LO IC₅₀ of greater than 10 μM. Accordingly, its 15-LO to5-LO ratio of activities is greater than 10, thus making the compound aspecific 15-LO inhibitor according to this invention.

All that is required to practice this invention is to administer to amammal an effective amount of any compound that is a 15-LO inhibitor.For example, the compounds of Formula I are useful for treatingatherosclerosis and inflammation by virtue of their ability to inhibit15-LO as established by the data in Table 1. Accordingly, any compoundthat is determined to inhibit 15-LO in a test system, such as describedabove, can be employed in this invention.

For use according to this invention, the compounds can be formulatedinto compositions suitable for administering to animals, includinghumans, for treating and preventing inflammation and atherosclerosis.The compounds can be formulated for administration by any route, forinstance orally, parenterally, topically, and rectally. For oraladministration, for example, a 15-LO inhibitor can be mixed with aninert diluent or with an assimilable edible carrier, or it may beenclosed in a hard or soft shell gelatin capsule, or it may becompressed into tablets, or it may be incorporated directly with thefood of the diet. For oral therapeutic administration, the activecompound may be incorporated with excipients and used in the form ofingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. Such compositions andpreparations should contain at least 1% by weight of active compound.The percentage of the compositions and preparations may, of course, bevaried and may conveniently be between about 5% to about 80% of theweight of the unit. The amount of active compound in suchtherapeutically useful compositions is such that a therapeuticallyeffective dosage will be obtained. Preferred compositions orpreparations according to the present invention are prepared so that anoral dosage unit form contains between about 5 and 1000 mg of activecompound, and ideally about 25 to about 750 mg.

The tablets, troches, pills, capsules, and the like may also containcommon pharmaceutical excipients such as binders, sweeteners, and thelike. Typical binders include gum tragacanth, acacia, corn starch, andgelatin, as well as excipients such as dicalcium phosphate. Typicaldisintegrating agents include corn starch, potato starch, alginic acid,and the like. A commonly used lubricant is magnesium stearate. Typicalsweetening agents are sucrose, lactose, or saccharin, and flavoringagents such as peppermint, oil of wintergreen, or cherry flavoring canbe utilized. When the dosage unit form is a capsule, it may contain, inaddition to materials of the above type, a liquid carrier. Various othermaterials may be present as coatings or to otherwise modify the physicalform of the dosage unit. For instance, tablets, pills, or capsules maybe coated with shellac, sugar, or both. A syrup or elixir may containthe active compound, sucrose as a sweetening agent, methyl andpropylparabens as preservatives, a dye, and flavoring such as cherry ororange flavor. Of course, any material used in preparing any dosage unitform should be pharmaceutically pure and substantially nontoxic in theamounts employed.

The 15-LO inhibitors can also be formulated for topical administration,for instance as patches, salves, creams, ointments, and the like. Agentscommonly utilized to enhance transdermal passage can also be employed.The compounds can also be formulated with waxes and the like forconvenient rectal administration.

The active 15-LO inhibitor may also be administered parenterally orintraperitoneally. Dispersions can also be prepared in glycerol, liquidpolyethylene glycols, and mixtures thereof and in oils. Under ordinaryconditions of storage and use, these preparations may contain apreservative to prevent the growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersions. In all cases, the form must be sterile andmust be fluid to the extent that easy syringability exists. It must bestable under the conditions of manufacture and storage and must bepreserved against the contaminating action of microorganisms such asbacteria and fungi. The carrier can be a solvent or dispersion mediumcontaining, for example, water, ethanol, a polyol (for example,glycerol, propylene glycol, and liquid polyethylene glycol, and thelike), suitable mixtures thereof, and vegetable oils. The properfluidity can be maintained, for example, by the use of a coating such aslecithin; by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. The prevention of theaction of microorganisms can be brought about by various antibacterialand antifungal agents, for example, parabens, chlorobutanol, phenol,sorbic acid, thimerosal, and the like. In many cases, it will bepreferable to include isotonic agents, for example, sugars or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompound in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredient into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum drying and the freeze-dryingtechnique which yield a powder of the active ingredient plus anyadditional desired ingredient from previously sterile-filtered solutionthereof.

As used herein, "pharmaceutically acceptable carrier" includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like. The useof such media and agents for pharmaceutical active substances is wellknown in the art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions.

It is especially advantageous to formulate parenteral compositions indosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the mammalian subjects to be treated; eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. The specification for the dosage unitforms are dictated by and directly dependent on (a) the uniquecharacteristics of the active material and the particular therapeuticeffect to be achieved, and (b) the limitations inherent in the art ofcompounding such an active material for the treatment of disease inliving subjects having a diseased condition in which bodily health isimpaired as herein disclosed in detail.

The principal active ingredient is compounded for convenient andeffective administration in effective amounts with a suitablepharmaceutically acceptable carrier in dosage unit form as hereinbeforedisclosed. The term "effective amount" means that quantity of a 15-LOinhibitor which has a positive therapeutic effect for treating orpreventing the inflammation or the atherosclerosis which affects themammal. Such amount is that which inhibits the 15-LO enzyme; in otherwords, a 15-LO inhibiting amount. A unit dosage form can, for example,contain the principal active compound in amounts ranging from about 5 toabout 1000 mg, with from about 25 to about 750 mg being preferred. Atypical dose will be about 50 to about 500 mg. In the case ofcompositions containing supplementary active ingredients, the dosagesare determined by reference to the usual dose and manner ofadministration of the said ingredients. The unit dosages typically willbe administered from one to four times per day, or as otherwise neededto effect treatment of the disease state.

The invention therefore is a method for treating and preventinginflammation and atherosclerosis in mammals. The compounds are effectivein inhibiting the activity of 15-LO, and as such can be administered toa mammal, including a human, to effectively diminish and treatatherosclerosis and inflammation. The compounds will be administered ata dose which is effective to treat atherosclerosis, typically from about1.0 to about 100 mg/kg of body weight of the subject being treated.

The compounds also are useful for treating and preventing inflammation,for example, swelling due to injuries, swelling around bones and joints,and the like. The compounds will be administered to an animal sufferingfrom inflammation in an anti-inflammatory effective amount that iseffective to treat the inflammation. Typical doses will be from about1.0 to about 100 mg/kg of body weight.

We claim:
 1. A method for treating or preventing inflammation oratherosclerosis in a mammal comprising administering a 15-LO inhibitingamount of a compound of Formula I ##STR38## wherein: R¹ is hydrogen orC₁ -C₆ alkyl;R², R³, R⁴, and R⁵ independently are hydrogen, C₁ -C₆alkyl, nitro, halo, CN, OR⁶, NR⁶ R⁷, --CO₂ R⁶, CONR⁶ R⁷, CH₂ OR⁶, or--CH₂ NR⁶ R⁷, or R² and R³, and R⁴ and R⁵, when attached to adjacentring atoms, can be --(CH₂)₃ or --(CH₂)₄ ; in which R⁶ and R⁷independently are hydrogen, C₁ -C₆ alkyl, phenyl, or benzyl, or whentaken together with the nitrogen to which they are attached, R⁶ and R⁷can complete a cyclic ring having from 3 to 7 carbon atoms; ##STR39## inwhich R⁸ and R^(8') independently are hydrogen or C₁ -C₆ alkyl, n is 0,1, or 2, and Z.sup.⊖ is an anion, or a pharmaceutically acceptable saltthereof.
 2. A method according to claim 1 employing a compound of theformula ##STR40## wherein R¹, R², R³, R⁴, R⁵, R⁸, and Z.sup.⊖ are asdefined above.
 3. A method of claim 2 employing a compound whereinZ.sup.⊖ is ClO₄.sup.⊖ or halo.
 4. A method of claim 3 employing acompound wherein R⁸ is hydrogen.
 5. A method of claim 4 employing acompound wherein R² and R³ both are hydrogen.
 6. A method of claim 5employing a compound wherein R⁵ is hydrogen.
 7. The method of claim 6employing a compound wherein R⁴ is NO₂.
 8. The method of claim 6employing a compound wherein R⁴ is CH₃.
 9. A method of claim 2 employinga compound wherein R², R⁴, and R⁵ are hydrogen and R³ is CH₃ O--.
 10. Amethod according to claim 1 employing a compound having the formula##STR41## wherein R¹, R², R³, R⁴, R⁵, and R⁸ are as defined above.
 11. Amethod according to claim 10 employing a compound wherein R¹, R⁴, R⁵,and R⁸ are hydrogen, R² and R³ independently are hydrogen or CH₃ O--.